J. J. Van Der Sluis

The localisation of treponemes and characterisation of the inflammatory infiltrate in skin biopsies from patients with primary or secondary syphilis, or early infectious yaws

OBJECTIVE--To study the localisation of treponemes and to analyse the inflammatory infiltrate in biopsy specimens from patients with primary or secondary syphilis, or early infectious yaws. MATERIALS AND METHODS--Skin biopsies originating from human lesions of primary (29x) or secondary (15x) syphilis (Rotterdam), or early yaws (18x) (West Sumatra) were studied. Different histochemical and immunohistochemical detection methods were used in this study. RESULTS AND CONCLUSION--The histochemical silver staining method according to Steiner revealed the presence of T. pallidum in all cases of primary syphilis studied. In 10 out of 14 cases of secondary syphilis, treponemes were demonstrated. With an immunofluorescence staining technique (IF) using anti-T. pallidum antiserum raised in rabbits (a-Tp), T. pallidum was demonstrated in 28 out of 29 cases of primary syphilis, and in 14 out of 14 studied cases of secondary syphilis. The silver staining method and IF showed identical localisations of T. pallidum (mainly in the dermal-epidermal junction zone or throughout the dermis). Using a-Tp antiserum in the indirect immunofluorescence technique, T. pertenue could be demonstrated in the dermis more often than with Steiner silver staining. However, epidermotropism of T. pertenue in yaws specimens was remarkable, compared with more mesodermotropism of T. pallidum; numbers of T. pertenue in the dermis were limited in all specimens. The dermal inflammatory infiltrate in primary and secondary syphilis was composed mainly of lymphocytes and plasma cells. In most cases more T (CD3 positive) cells than B (CD22 positive) cells were present. Regarding T cell subpopulations, in primary syphilis, T helper/inducer (CD4 positive) cells predominated in 86% of cases. In secondary syphilitic lesions, numbers of T helper/inducer cells were less frequent than or equal to T-suppressor/cytotoxic (CD8 positive) cells in 60% of cases. Remarkably, in yaws specimens the inflammatory infiltrate consisted mainly of IgG, but also IgA and IgM producing plasma cells. T or B lymphocytes were scarce, which is in sharp contrast with findings in syphilitic lesions.

Primary and secondary syphilis: a histopathological study.

We present a study of biopsies taken from skin lesions of 44 patients presenting with primary or secondary syphilis. In most primary lesions erosion or, more often, ulceration was present, with a dense inflammatory infiltrate. In secondary syphilis a wide variety of histological changes was present. Blood vessels were frequently involved, with marked endothelial swelling and often proliferation. Treponemes were demonstrated with the Steiner staining method in all investigated cases of primary syphilis and in 71% of secondary syphilis cases. Treponemes were present throughout the dermis, particularly perivascularly, and in the dermal–epidermal junction zone. In two specimens of secondary syphilis treponemes were located predominantly in the epidermis, but there were always some microorganisms demonstrable in the dermis. The inflammatory infiltrate was often located in a perivascular coat-sleeve-like arrangement. In this study plasma cells and lymphocytes were present in all specimens of primary and secondary syphilis. Syphilitic lesions differed from yaws lesions mostly in the location of treponemes and the affection of blood vessels. In this histopathological study of early syphilis, treponemes did not show the epidermiotropic character of yaws, and blood vessel changes were more pronounced than in yaws. Unfortunately, due to the protean histopathological manifestations described in venereal syphilis and in yaws, these two treponemal diseases cannot always be differentiated on histological grounds alone.

Early yaws : a light microscopic study

This paper presents the light microscopic findings in biopsies of skin lesions from 45 patients, in whom a diagnosis of early yaws was suspected. In 27 cases typical light-microscopic features of yaws were observed, consisting of parakeratosis or crust containing exudate, marked acanthosis with widening and elongation of the rete ridges or pseudo-carcinomatous hyperplasia and spongiosis. Intraepidermal microabscesses consisting of polymorphonuclear leucocytes were frequently encountered. In a large majority a moderate to dense infiltrate was present, composed mainly of lymphocytes and plasma cells. Vascular changes consisted of only slight endothelial cell proliferation and thickening of vessel walls. Steiner staining revealed the presence of treponemes in the epidermis in 23 of 27 cases. Remarkably, clusters of treponemes were also seen in the papillary dermis in three out of 23 cases. Seven other cases were strongly suggestive of yaws. Other histopathological diagnoses were made in 6 patients, due to the simultaneous occurrence of other skin diseases. The remaining five specimens did not contain enough tissue to allow conclusions to be made.

Yaws in West Sumatra, Indonesia: Clinical manifestations, serological findings and characterisation of new Treponema isolates by DNA probes

The results of a yaws survey on the island of Sumatra in Indonesia are presented. The prevalence of yaws in the investigated region was found to be very high, a minimum of 300 cases per 100,000 individuals, which indicates that yaws is far from being eradicated and that campaigns for treatment are necessary. Patients suffering from early infectious yaws showed florid skin lesions. Of 101 serum samples from such patients, 100 had a positive reaction in one or more treponemal tests. TheTreponema pallidum haemagglutination assay was found to be the most sensitive test (97 % positive) in detecting antibodies againstTreponema pallidum subsp.pertenue, followed by the fluorescent treponemal antibody absorption test (94 %), the Venereal Disease Research Laboratory test and the TmpA enzyme immunoassay (91 %), and analysis by Western blot usingTreponema pallidum antigens (88 %). Of 42 asymptomatic contacts of yaws patients 32 showed positive reactions in one or more tests, indicating that many people in the investigated region have been infected with treponemes. Eight newTreponema pallidum subsp.pertenue strains were isolated from yaws skin lesions. In vitro amplification of treponemal DNA and hybridisation with specific DNA probes showed that all eight strains were identical withTreponema pallidum subsp.pertenue CDC 2575, with regard to the subsp.pertenue specifictyfl gene.

An IgM capture enzyme linked immunosorbent assay to detect IgM antibodies to treponemes in patients with syphilis.

A new IgM capture enzyme linked immunosorbent assay (ELISA) was compared with the 19S(IgM) fluorescent treponemal antibody absorption (19S(IgM)FTA-ABS) test for detecting IgM antibodies to treponemes. Serum samples from 180 people, 109 with various stages of untreated syphilis, 45 with treated syphilis, and 26 non-infected, were investigated. In all diagnostic groups of syphilis the reactivity of the IgM capture ELISA was similar to that of the 19S(IgM)FTA-ABS test except in untreated neurosyphilis, for which the IgM capture ELISA was significantly less sensitive. The IgM capture ELISA was very sensitive in congenital (100%, 5/5) and primary (82%, 18/22) syphilis, but less sensitive in secondary (60%, 12/20), latent (53%, 16/30), neurosyphilis (34%, 11/32), and treated (11%, 5/45) syphilis. False positive IgM capture ELISA results were not found in five people who gave false positive Venereal Disease Research Laboratory (VDRL) reactions or in 21 neonates born to mothers adequately treated for syphilis before or during pregnancy. This indicated that the IgM capture ELISA was very specific. The course of antitreponemal IgM reactivity after treatment of early infectious syphilis was followed up in six patients. The quantity of IgM antibody declined in nearly all patients after treatment, but still remained detectable in five patients up to six months after treatment. In contrast, non-treponemal antibodies measured by the VDRL test disappeared in four out of six patients within five months from starting treatment. In conclusion, the IgM capture ELISA may be useful for easy and sensitive detection of IgM antibodies to treponemes in patients with congenital and primary syphilis. A positive test result in these cases indicates that patients should receive treatment if they have not been treated recently. The test is not, however, recommended to replace the VDRL test to monitor patients treated for syphilis.

Laboratory techniques in the diagnosis of syphilis : a review

Introduction At present, the laboratory diagnosis of syphilis is based on the visualisation of the pathogen, Treponema pallidum and/or the demonstration of specific antibodies in body fluids by means of serological tests. In this article the current methods, the test procedures that have only recently been introduced to diagnose syphilis and the diagnosis based on molecular biology techniques are reviewed.

Radiological and dermatological findings in two patients suffering from early yaws in Indonesia.

Two children suffering from early yaws in Indonesia are presented. Apart from skin lesions and a positive treponemal serology in both patients, involvement of tubular bones, particularly of the hands, was revealed by radiological examination. In one patient involvement of a distal phalanx was remarkable. Early diagnosis and treatment of yaws is important since a delay in treatment may result in severe and irreversible bone deformities of the late stage of the disease. This report illustrates that radiological changes, although rare, can still be detected in the early stage of yaws in areas where yaws is resurgent nowadays.

Enzyme-linked immunofiltration assay for rapid serodiagnosis of syphilis

A new rapid technique for detection of serum treponemal antibodies is described which is based on an enzyme-linked immunoassay using nitrocellulose as solid phase. With this technique antigen-antibody binding is accelerated by the filtration of the antibody solution through the antigen-coated nitrocellulose filter instead of its remaining over the solid phase for incubation. Test results are available in less than 15 min. Serum specimens from 255 syphilitics and 829 non-infected subjects were investigated. The sensitivity and specificity of theTreponema pallidumenzyme-linked immunofiltration assay were comparable to those of theTreponema pallidumhaemagglutination assay and the fluorescent treponemal antibody absorption test.

Ultrastructural aspects of infection with Treponema pallidum subspecies pertenue (Pariaman strain).

OBJECTIVE--To study ultrastructural aspects of infection with Treponema pertenue (Pariaman strain), originating from West Sumatra, Indonesia. MATERIALS AND METHODS--Biopsy material originating from skin lesions in ten young children suffering from early infectious yaws in Indonesia, and rabbit testicular tissue inoculated with T pertenue. Human skin as well as rabbit testicular tissue was examined by means of conventional electron microscopy. RESULTS--In human skin, treponemes were found in interepidermal spaces in 5 out of 10 specimens. In two of five positive specimens, treponemes were also seen in the dermis. In one out of five specimens from rabbit testicular tissue a profusion of treponemes was found lying in the interstitial myxomatous tissue. Microorganisms showed no adhesion to fibroblasts. CONCLUSION--This ultrastructural study of T pertenue demonstrated the scarcity and focal distribution of treponemes in tissue and did not reveal any morphological differences from the Gauthier strain of T pertenue. No differences from the ultrastructure of T pallidum were observed either.

Mucopolysaccharides in suspensions of Treponema pallidum extracted from infected rabbit testes.

The amount and nature of mucopolysaccharides present in extraction fluids routinely obtained in the isolation procedure of Treponema pallidum from infected rabbit testes was investigated. The mean quantity of mucopolysaccharides extracted from both testes of groups of 10 rabbits was 3.09 mg after infection for seven days and 26.88 mg after infection for 12 days, while from the testes of uninfected rabbits a mean of 0.42 mg was obtained. On electrophoresis the isolated mucopolysaccharides showed only one single band with the migration characteristics of hyaluronic acid. This band disappeared completely after pretreatment with hyaluronidase from bovine testes, which showed that during infection with T pallidum increasing amounts of hyaluronic acid accumulate. They can, at least in part, be extracted by a gentle extraction procedure, suggesting that this material binds loosely. The amount of hyaluronic acid isolated 12 days after infection showed positive correlations with the wet weight of testes as well as the number of treponemes isolated; seven days after infection such correlations were not present.